Rapid Plasma Proteome Profiling via Nanoparticle Protein Corona and Direct Infusion Mass Spectrometry

Noninvasive
detection of protein biomarkers in plasma is crucial
for clinical purposes. Liquid chromatography–mass spectrometry
(LC–MS) is the gold standard technique for plasma proteome
analysis, but despite recent advances, it remains limited by throughput,
cost, and coverage. Here, we introduce a new hybrid method that integrates
direct infusion shotgun proteome analysis (DISPA) with nanoparticle
(NP) protein corona enrichment for high-throughput and efficient plasma
proteomic profiling. We realized over 280 protein identifications
in 1.4 min collection time, which enables a potential throughput of
approximately 1000 samples daily. The identified proteins are involved
in valuable pathways, and 44 of the proteins are FDA-approved biomarkers.
The robustness and quantitative accuracy of this method were evaluated
across multiple NPs and concentrations with a mean coefficient of
variation of 17%. Moreover, different protein corona profiles were
observed among various NPs based on their distinct surface modifications,
and all NP protein profiles exhibited deeper coverage and better quantification
than neat plasma. Our streamlined workflow merges coverage and throughput
with precise quantification, leveraging both DISPA and NP protein
corona enrichment. This underscores the significant potential of DISPA
when paired with NP sample preparation techniques for plasma proteome
studies.

血浆中蛋白质生物标志物的无创检测对于临床应用至关重要。液相色谱-质谱联用法（liquid chromatography–mass spectrometry, LC–MS）是血浆蛋白质组分析的金标准技术，但尽管近年来技术取得了长足进步，该方法仍受限于通量、成本与覆盖范围。本研究提出一种新型混合分析方法，将直接注入式鸟枪法蛋白质组分析（direct infusion shotgun proteome analysis, DISPA）与纳米颗粒（nanoparticle, NP）蛋白质冠富集技术相结合，以实现高通量、高效的血浆蛋白质组学分析。该方法仅需1.4分钟的采集时长即可实现超过280种蛋白质的鉴定，单日潜在样本处理通量可达约1000份。所鉴定的蛋白质参与了多条具有重要研究价值的通路，其中44种为美国食品药品监督管理局（Food and Drug Administration, FDA）批准的生物标志物。本研究针对多种纳米颗粒与不同浓度梯度，对该方法的稳健性与定量准确性进行了评估，其平均变异系数为17%。此外，由于不同纳米颗粒的表面修饰特性存在差异，其蛋白质冠谱特征也各不相同；且相较于纯血浆样本，所有纳米颗粒的蛋白质谱均具备更深的覆盖度与更优异的定量性能。本研究的简化流程整合了DISPA与纳米颗粒蛋白质冠富集技术，在保证覆盖度与通量的同时实现了精准定量。这充分彰显了将DISPA与纳米颗粒样本制备技术相结合应用于血浆蛋白质组研究的巨大潜力。